Renal-Electrolyte Division, Department of Medicine, University of Pittsburgh School of Medicine.
Renal-Electrolyte Division, Department of Medicine, University of Pittsburgh School of Medicine;
J Vis Exp. 2022 Oct 6(188). doi: 10.3791/64584.
In addition to forming a high-resistance barrier, the urothelium lining the renal pelvis, ureters, bladder, and proximal urethra is hypothesized to sense and transmit information about its environment to the underlying tissues, promoting voiding function and behavior. Disruption of the urothelial barrier, or its sensory/transducer function, can lead to disease. Studying these complex events is hampered by lack of simple strategies to alter gene and protein expression in the urothelium. Methods are described here that allow investigators to generate large amounts of high-titer adenovirus, which can then be used to transduce rodent urothelium with high efficiency, and in a relatively straightforward manner. Both cDNAs and small interfering RNAs can be expressed using adenoviral transduction, and the impact of transgene expression on urothelial function can be assessed 12 h to several days later. These methods have broad applicability to studies of normal and abnormal urothelial biology using mouse or rat animal models.
除了形成高电阻屏障外,肾盂、输尿管、膀胱和尿道近端的尿路上皮还被假设能够感知其环境并将信息传递到下面的组织,从而促进排尿功能和行为。尿路上皮屏障或其感觉/换能器功能的破坏可导致疾病。由于缺乏改变尿路上皮基因和蛋白表达的简单策略,研究这些复杂事件受到了阻碍。本文描述了可使研究人员生成大量高滴度腺病毒的方法,然后可使用这些腺病毒以高效且相对直接的方式转导啮齿动物的尿路上皮。可以使用腺病毒转导来表达 cDNA 和小干扰 RNA,并且可以在 12 小时到几天之后评估转基因表达对尿路上皮功能的影响。这些方法广泛适用于使用小鼠或大鼠动物模型研究正常和异常尿路上皮生物学。